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In Vitro Dynamics of Kinesin 1 and Myosin VI

驱动蛋白 1 和肌球蛋白 VI 的体外动力学

基本信息

批准号：
8217276
负责人：
PAUL R SELVIN
金额：
$ 34.48万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-06-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8217276
关键词：
ATP phosphohydrolase Address Affect Alzheimer&apos s Disease Amino Acids Antineoplastic Agents Binding CENP-E protein Calmodulin Cells Central Nervous System Diseases Chimera organism Color Cysteine Data Defect Development Dimensions Dimerization Family member Fluorescence Grant Hand Head Head and neck structure Health Heart Diseases Image In Vitro Kinesin Kinetics Length Leucine Zippers Light Link Maintenance Malignant neoplasm of brain Measures Microtubules Molecular Motors Motion Motor Motor Activity Movement Myosin ATPase Neck Nucleotides Occupations Organelles Paper Pharmaceutical Preparations Phase Physiological Play Positioning Attribute Probability Protein Region Proteins Regulation Resolution Rice Role Running SWI1 Science Seminal Signal Transduction Structure Tail Techniques Testing Time Torsion Walking Work arm cancer type cell motility crosslink deafness falls fluorescence imaging fluorophore graduate student millisecond mutant myosin VI nanometer optical traps single molecule single-molecule FRET small molecule tool

项目摘要

DESCRIPTION (provided by applicant): Molecular motors--kinesin and myosins--play a crucial role in the maintenance and development of the organization, motility, and signaling of healthy cells. Central nervous-system disorders, such as Alzheimer's disease and certain types of cancer, all arise from molecular motors gone awry. There are, however, several fundamental questions about how kinesin-1 moves. (Kinesin-1 is the "standard-bearer" of kinesins.) Also largely unknown is how Myosin VI moves. To answer these questions we will address the motors at the in vitro, single molecule level. We will apply a number of single molecule tools, including some that have been developed in the first 4 years of this proposal. We will use FIONA (Fluorescence Imaging with One Nanometer Accuracy), DOPI (Defocused Orientation and Position Imaging), and SHREC (Single Molecule High REsolution Colocalization). The theme of the kinesin part is: how is motility affected by each of kinesin's parts, including the head, the coiled-coil, and the tail? The theme of the myosin VI part is: how does such a small motor take such a large step? We will present unpublished data which suggests that kinesin is bound by both heads during a run, making the probability of falling off very low. We will also suggest that the large non-helical region in kinesin-1's coiled-coiled region allows this motor to walk in an "asymmetric fashion," allowing the cargo to point forward. Finally, we will suggest that full-length kinesin takes "pauses" due to the tail-region folding over and interacting with the head region and possibly the microtubule. With regards to myosin VI, we have an enormous amount of preliminary data. It will suggest that the head undergoes a 180¿ swing during the powerstroke. Furthermore, we suggest that a 3-helix motif in the lever arm uncoils and creates an unprecedented 24 nm extension, which allows the motor to take a 36 nm step. PUBLIC HEALTH RELEVANCE Molecular motors have the job of moving and organizing organelles within a cell. Problems within the motors cause brain cancer, Alzheimer's disease, etc. We seek to understand the basic workings of kinesins and myosin VI, two important motors.

描述（由申请人提供）：分子马达-驱动蛋白和肌球蛋白-在健康细胞的组织、运动和信号传导的维持和发展中起关键作用。中枢神经系统疾病，如阿尔茨海默病和某些类型的癌症，都是由分子马达出了问题引起的。然而，关于驱动蛋白-1如何移动，还有几个基本问题。（驱动蛋白-1是驱动蛋白的“旗手”。）同样未知的是肌球蛋白VI如何移动。为了回答这些问题，我们将在体外，单分子水平上解决电机。我们将应用一些单分子工具，包括在本提案的前4年中开发的一些工具。我们将使用FIONA（一纳米精度的荧光成像），DOPI（散焦方向和位置成像）和SHREC（单分子高分辨率共定位）。驱动蛋白部分的主题是：驱动蛋白的每个部分，包括头部，卷曲螺旋和尾部，是如何影响运动的？肌球蛋白VI部分的主题是：如此小的马达如何迈出如此大的一步？我们将提供未发表的数据，这些数据表明驱动蛋白在运行期间被两个头部结合，使得脱落的概率非常低。我们还将提出，驱动蛋白-1的卷曲-卷曲区域中的大的非螺旋区域允许该马达以“不对称的方式”行走，允许货物指向前方。最后，我们将建议，全长驱动蛋白采取“暂停”由于尾部区域折叠和相互作用的头部区域，并可能微管。关于肌球蛋白VI，我们有大量的初步数据。这将表明，头部经历了180 <$摆动在动力冲程。此外，我们认为杠杆臂中的3-螺旋基序展开并产生前所未有的24 nm延伸，这使得电机可以采取36 nm的步骤。公共卫生相关性分子马达的工作是移动和组织细胞内的细胞器。在电机的问题导致脑癌，阿尔茨海默氏症等，我们试图了解驱动蛋白和肌球蛋白VI，两个重要的电机的基本工作。